Acrylamide levels determination by a colorimetric method based on biotechnological enzymes

INTRODUCTION Acrylamide is a carcinogenic and mutagenic compound found in many industrially processed foods and beverages, including coffee. It has been labelled by the International Agency for Research on Cancer (IARC) as a probable human carcinogen;1 very recently, it has been classified as a carcinogen and mutagen by the European Union.2 In 2002, researchers from Sweden reported finding acrylamide in certain baked and fried foods such as potato crisps and French fries. The general findings are that acrylamide will likely be found in foods containing both sugars and proteins heated by means other than boiling, since generation of acrylamide and methylacrylamide is mainly the result of Maillard reaction between the amino acid asparagine and reducing sugars. Until now, the methods applied for quantitation of acrylamide have been GC-MS and HPLC-MS. MATERIALS AND METHODS The aim of the present work is the set up and validation of an enzymatic test kit, which allows quantitation of acrylamide, after some preliminary operations involving proprietary, thermo-stable engineered enzymes, through the use of a colorimeter. The proposed method is much simpler and more economical than the classical chromatography-MS approach. The enzymatic test kit has been used for acrylamide quantitation in some coffees; the same samples have been analyzed by a standard HPLC-MS method, for the validation of the enzymatic method. RESULTS AND DISCUSSION Results showed that there is a close correspondence between the two methods and that the kit provides a cost-effective method for the determination of acrylamide levels in food. Furthermore, in the analyzed food samples the acrylamide level showed to be quite low, with concentrations in the range of 88-250 ppb.